Cloning and expression of squalene epoxidase from the pathogenic yeast Candida albicans
Details
Serval ID
serval:BIB_0504EEBA594F
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Cloning and expression of squalene epoxidase from the pathogenic yeast Candida albicans
Journal
Gene
ISSN
0378-1119 (Print)
Publication state
Published
Issued date
04/1997
Volume
189
Number
1
Pages
119-26
Notes
Comparative Study
Journal Article --- Old month value: Apr 11
Journal Article --- Old month value: Apr 11
Abstract
The allylamine antimycotic terbinafine prevents the formation of sterols by specifically inhibiting squalene epoxidase (SE). The biological and biochemical action of terbinafine on fungal pathogens has been well investigated, but little is known at the molecular level. Here we report the cloning, sequencing and expression of the target of terbinafine from the major pathogen Candida albicans. A C. albicans genomic DNA library was constructed in gamma ZAP Express and screened with a DNA fragment obtained by polymerase chain reaction with two primers designed from sequences common to Saccharomyces cerevisiae and rodent SEs. Two types of clone, approximately 3.9 kbp and 4.1 kbp, were isolated. Both contained an identical open reading frame of 1488 nucleotides, while a few sequence differences were found in the flanking regions, suggesting an allelic heterogeneity. The deduced protein sequence of C. albicans SE, 496 amino acids (55324 Da), is 54% and 41% identical to those of S. cerevisiae and rat, respectively. A 1.8-kb transcript was observed on Northern blots of C. albicans mRNA. Polyclonal antibodies, raised against an internal peptide of C. albicans SE, recognized a protein associated with the particulate fraction of M(r) 55000 on Western blots of C. albicans extracts. C. albicans SE was overexpressed in S. cerevisiae with the expression vector pYES2. In homogenates from S. cerevisiae overexpressing the C. albicans protein SE activity was 10-fold higher than the endogenous activity from controls.
Keywords
Amino Acid Sequence
Animals
Base Sequence
Candida albicans/*enzymology/*genetics/pathogenicity
Cloning, Molecular
Fungal Proteins/*genetics/isolation & purification
Gene Expression Regulation, Fungal
Genetic Vectors/chemistry
Immunoblotting
Mice
Molecular Sequence Data
Oxygenases/*genetics/isolation & purification
Rats
Saccharomyces cerevisiae/genetics
Sequence Alignment
Squalene Monooxygenase
Transcription, Genetic
Pubmed
Web of science
Create date
25/01/2008 16:32
Last modification date
20/08/2019 12:26